Preserving Pharmaceutical Products: Patrick Crowley

Pharmaceutical Products Modes of administration vary Oral Injection or infusion (parenteral) Inhalation (oral Intranasal Applied to skin ( topical ) - creams/ointments/lotions) may be applied to mucosal surfaces may be applied to damaged tissue Ophthalmic Products Generally have long shelf lives 3-5 years in many cases Some product components possess intrinsic antimicrobial activity. eg antimicrobial drugs Other product components may confer or aid preservation

Preservatives in Prescription Products ("drugs@fda".com) Preservative Oral Topical Ophthalmic Parenteral p- hydroxybenzoate esters ("Parabens") 39 14 None None Parabens/Na benzoate 4 13 None None Na Benzoate 24 2 (antifungals) None 1 Sorbic acid/potassium Sorbate 5 2 (one with methyl paraben) 1 None Benzalkonium Chloride 1 1 31 None Benzodecinium Bromide None None 2 None Benzyl alcohol 1 9 None 3 Phenylethyl alcohol None 2 None None Phenoxy ethanol None 4 None None m-cresol None None None 1 Stearalkonium Bromide None 2 None None Chlorbutanol None None None 1 Thimoresal None None 1 1

70 Product Forms in US (Y axis = percentage) (Oral and Parenteral only) Oral dosage 70 Parenteral dosage 60 60 50 50 40 40 30 30 20 20 10 10 0 0 Solution Lyo for recon Suspension Solid for recon Lyo for susp Emulsion

Products requiring Preservation Product Type Preservative? Solids for oral dosage eg tablets/capsules no Liquids (aqueous) for oral dosage multidose (incl when constituted from solid on dispensing) yes single dose (eg in a sachet or dispersible/effervescent tablet) no Sterile liquids (aqueous) for infusion/injection multidose yes single-dose no Ophthalmic Preparations aqueous multidose yes single dose no if sterile Topical Products (applied to skin, mucosal surfaces etc) ointments (non-aqueous) no creams/lotions: (aqueous): multidose yes single-dose no if sterile Liquids for nasal inhalation (aqueous, multidose) yes

Preservatives in Pharmaceuticals Preservative must be suitable/appropriate for the mode of dosage/use compatible with the drug and other ingredients effective throughout the shelf life/usage period of the product (preservative stability) Inclusion level must be minimal for antimicrobial efficacy Regulatory and pharmacopeial guidelines - the minimum concentration be used to give the required level of efficacy (EMEA) - - below a level that may be toxic to human beings (USP) A preservative must not be a substitute for poor GMP

Exclusion of a Preservative: Justifications Active ingredient(s) provide(s) the requisite effect antimicrobial agent(s) in oral, parenteral, topical etc products must provide antibacterial and antifungal effect meet pharmacopeial Antimicrobial Efficacy Test requirements Other product component(s) has/have antimicrobial effects sucrose in oral products non-aqueous solvents in topical products glycerol, propylene glycol, ethanol lower the water activity

Antimicrobial Efficacy Testing Detailed in Pharmacopeias (USP, Ph.Eur, BP J.P copeia. Test organisms comprise common bacteria and molds/fungi Bacteria P.aeruginosa and S.aureus (+ E.coli in USP) Molds/fungi C.albicans and A.brasiliensis Additional organisms may be included where appropriate e.g possible contaminants in facility, materials, operators etc E.coli in oral products Z.rouxii in sucrose-containing products Performance standards include microcidal and/or microstatic activity depending on mode of product use Testing procedures are (mostly) common Some differences in performance standards USP/JP versus Ph.Eur/BP Less stringent requirements for antacid products in USP and JP.

Performance Requirements Required Log 10 Reduction (minimum) Reference Product Type (aqueous) Organisms * 6 24 48 7 14 28-days hours hours hours days days A 2 3 No recovery Bacteria Parenteral and B 1 3 No increase from day 7 Ophthalmic A 2 Fungi B 1 Ph.Eur and British P'copeias USP and Japan P'copeias Oral Topical Bacteria 3 Fungi 1 Bacteria Fungi A 2 3 No increase from day 7 B 3 No increase from day A 2 14 B 1 * "A" is recommended. "B" criteria may be acceptable if adverse reactions are a (justified) issue. Parenterals, sterile nasal/ophthalmic products (aqueous) Oral, except antacids Antacids (aqueous) Non-sterile topical, nasal, aural (aqueous) No increase from day Bacteria 1 3 14 Fungi No increase from initial No increase from day Bacteria 1 14. Fungi No increase from initial Bacteria Fungi Bacteria 2 Fungi No increase from day 14 No increase from initial No increase from day 14 No increase from initial

Preservative Performance Requirements Product meets pharmacopeial preservative performance standards Is compatible (chemical, physical) with the other product components Preservative is effective over the product ph range. Effective at the lower limit for preservative content in the product specification Effect is sustained throughout product lifetime (including use). Solubility in the product is adequate and not compromised by conditions encountered during product manufacture, storage, transport and use. Does not adversely affect patient-sensitive quality attributes such as taste, odour, irritation etc at the inclusion level in the product.

Preservative Efficacy and Product ph Preservative Active Moiety ph for optimum activity p- hydroxybenzoate esters ("parabens") Ester ph 4-8 Benzoic acid/salts Unionised (acid) <ph 4.5 Benzalkonium Cl Cation ph 4-10 Benzyl alcohol <ph 5.0 Chlorhexidine Cation ph 5-7 Propionic Acid Unionised (acid) ph 3.9 Sorbic acid/salts Unionised (acid) ph 4.5 Phenylmercuric salts Cation ph 5-8 Thimerosal "acidic ph"

ph, and Ionisation of Organic Acid Preservatives ph % not ionised (unionised) Benzoic acid Sorbic acid Propionic acid pka 4.2 4.76 4.88 2 99.4 99.8 99.9 3 94.1 98.4 98.7 4 61.3 85.2 88.4 5 13.70 36.5 43.50 5.5 4.78 15.4 19.23 6 1.56 5.4 7.04 6.5 0.50 1.8 2.34 7 0.16 0.6 0.76

Interactions with Excipients/Packaging Components Preservative Benzalkonium chloride Benzoic acid Benzyl alcohol Cetrimide Chlorbutanol Chlorhexidine Adsorbent/Substrate Hypromellose Filter Membranes Kaolin Polyethylene, Natural Rubber Bentonite Polyethylene Various polymeric excipients eg sodium carboxymethylcellulose p- hydroxybenzoate esters Ion Exchange Resins, some plastics Phenoxy ethanol Phenylmercuric salts Sorbic acid/sorbates Thimerosal PVC, Cellulose-based excipients Various suspending agents Polypropylene, PVC, Polyethylene Polyethylene, other plastics, rubber

Residues and Additives in Pharmaceutical Excipients Excipient Residue/Additive povidone, crospovidone, fixed oils, lipids polysorbates benzyl alcohol polyethylene glycol microcrystalline cellulose starch talc stearate salts hydroxypropylmethyl & ethyl celluloses peroxides antioxidants peroxides benzaldehyde aldehydes, peroxides, organic acids lignin, hemicelluloses, water formaldehyde heavy metals alkaline residues glyoxal

Preservatives Susceptible to Adsorption Compound Benzalkonium chloride Benzoic acid Benzyl alcohol Chlorhexidine Parabens Phenoxy ethanol Phenylmercuric salts Sorbic acid/sorbates Thimerosal Adsorbent PVC, Polyethylene Kaolin Polythylene, natural rubber Polymer-based excipients, Contact lense material Ion exchange resins, some plastics Cellulose-based excipients, PVC. Various suspending agents Polypropylene, PVC, Polyethylene Polyethylene

Oil/Water Partition Coefficients for Preservatives Preservative Oil Partition Coefficient Almond Oil 7.5 Methyl Paraben Mineral Oil 0.1 Isopropyl Myristate 18 Diethyl Adipate 200 Ethyl Paraben 26 Propyl Paraben 87 Soya Bean Oil Butyl Paraben 280 Benzoic Acid 6.1 Sorbic Acid Phenol Almond Oil Arachis Oil 3.3 5 Mineral Oil Mineral Oil 0.21 0.07

Preservative Enhancement Combinations of Preservatives Oral liquids Sucrose, glycerol, propylene glycol, (+ a formal preservative) Topicals Propylene glycol, ethanol, EDTA (+ a formal preservative) Parenterals EDTA (+ a formal preservative) Ophthalmics and (occasionally) intranasal EDTA with benzalkonium chloride (BAC/BKC) at inclusion levels of 0.0075% - 0.02%)

Combinations of Preservatives Preservative Preservation Capability Potential Companion Preservative Benzalkonium Chloride More active against g-positive bacteria. Benzyl alcohol and phenylethanol can enhance anti- Pseudomonas activity. EDTA facilitates reduced inclusion levels to reduce irritancy in ophthalmic products. Benzoic Acid/Na salt p- hydroxybenzoic acid esters ("Parabens") Benzyl alcohol Chlorhexidine Phenylethanol Phenoxyethanol Suitable for oral liquids with acidic ph values (not bitter). Activity is reduced as ph increases. Antifungal effect less susceptible to ph than antibacterial activity Particularly effective against Yeasts and Moulds. Less effective against Gram-negative organisms. Activity increases with increase in alkyl chain length. Effective against gram positive bacteria, yeasts, moulds; less active against gram- negative bacteria. Active against a wide range of bacteria, except for some Pseudomonas. Antifungal activity is limited. Good anti-pseudomonas activity so is often used to improve anti-pseudomonas activity of a companion preservative. Good anti-pseudomonas activity. Poor antifungal. Combinations with p-aminobenzoates can extend spectrum of activity. Combinations of two esters can overcome solubility constraints. Possible synergies with phenylethanol. Enhance the activities of benzalkonium chloride and chlorhexidine against pdeudomonas. Combination with Sorbate can enhance antifungal effect. Anti- Pseudomonas activity improved in combinations with benzalkonium chloride and chlorhexidiene gluconate. Synergistic effects with parabens against Yeasts and Molds.

Benzalkonium Chloride (BKC) in Ophthalmic Products Wide spectrum of antimicrobial activity Effective over wide ph range (ph 4-9). May enhance drug penetration to anterior chamber disrupts the hydrophobic barrier of corneal epithelium Effective at low inclusion levels Low allergenic potential but Long term use can cause allergic/inflammatory reactions/corneal damage Co-formulation with EDTA helps reduce inclusion levels of BKC

The Future What might change?

CAR-T Cell Therapy. Now (autologous therapy): patient-specific plasma, separate the T Cells incorporate the cancer-seeking protein (CD 19) using gene editing techniques culture the modified cells to increase numbers formulate the product test and ship to Center administer to patient. The Future??? (allogenic therapy) plasma from healthy donors rather than cancer-patient-specific plasma separation, gene-editing, formulation, shipping etc as above stockpile material(s) at the Treatment Center? administer to patient following diagnosis of cancer type